Tak1 CRISPR Activation Plasmid (m): sc-424044-ACT

Mouse Map3k7 encodes TAK1, a serine/threonine MAP3K that integrates signaling downstream of receptors for TNF, IL-1, TLR ligands, and TGF-β family members to coordinate inflammatory and stress responses. TAK1 phosphorylates MAP2Ks to engage JNK and p38 MAPK cascades and activates the IKK complex to drive NF-κB–dependent transcription, thereby shaping cytokine production, apoptosis, and cell survival decisions. Through these pathways, TAK1 influences innate immune activation, tissue homeostasis, and cellular responses to genotoxic and oxidative stress. Dysregulated TAK1 signaling has been implicated in inflammatory pathology, fibrosis-related processes, and oncogenic signaling networks, making Map3k7 a widely used node for mechanistic studies of pathway cross-talk.

Tak1 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Map3k7 expression without altering the underlying DNA sequence.

Tak1 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Map3k7 locus in human cell lines. The system is built around a catalytically inactive Cas9 (dCas9) carrying two inactivating mutations (D10A and N863A) that eliminate nuclease activity while preserving DNA binding. This dCas9 is fused to VP64, a potent transcriptional activator, and is co-expressed with a blasticidin resistance gene for selection. The second plasmid encodes the MS2-p65-HSF1 fusion protein, a secondary activator complex that works in concert with dCas9-VP64, alongside a hygromycin resistance gene. The third plasmid encodes a target-specific 20 nt sgRNA fused to two MS2 RNA aptamers that recruit the MS2-p65-HSF1 complex to the activation site, accompanied by a puromycin resistance gene. The three plasmids are delivered at a 1:1:1 mass ratio for balanced expression of all system components.

Once assembled at the target locus, the SAM complex binds within approximately 200 bp upstream of the Map3k7 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Tak1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Map3k7 locus and enabling the study of Tak1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Tak1 pathway restoration in tumor cells with silenced or reduced Map3k7 expression.

For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.